Aluminium (Al) toxicity in acid soils severely limits forage productivity, and dissecting Al-tolerance mechanisms is crucial for securing forage supply in acid-soil regions. White clover (Trifolium repens L.), an excellent acid-soil-adapted forage with pronounced Al tolerance, serves as an ideal model for studying legume Al-tolerance mechanisms. We exposed white clover seedlings to gradient Al³⁺ concentrations (0, 2, 4, 6, 10 mmol·L⁻¹) to characterize the core physiological and molecular responses underlying its Al adaptation. Al³⁺ stress inhibited growth in a concentration-dependent manner, suppressing both root and shoot development. Roots adopted a prioritised defence strategy, with enhanced antioxidant enzyme activity and soluble sugar accumulation mitigating oxidative damage. Transcriptome analysis revealed coordinated regulation of key pathways: flavonoid biosynthesis showed a core inhibition-branch-specific activation pattern, photosystem-related genes were upregulated to reinforce photosynthetic function, and hormone signalling networks were extensively rewired with divergent responses among auxin, gibberellin, cytokinin, ABA and JA pathways. White clover copes with Al³⁺ stress via an integrated mechanism featuring root-prioritised defence, photosynthetic maintenance and hormone network remodelling. These findings provide new insights into legume Al tolerance and a framework for breeding Al-tolerant forages. Future studies will quantify Al content in shoots and roots, and perform functional validation of upregulated hormone-related genes to clarify their roles in the Al-tolerance regulatory network.

25-Hydroxyvitamin D (25(OH)D), a metabolite of vitamin D, has demonstrated anticancer properties; however, the role of alternative splicing in mediating these effects remains poorly understood. In this study, we reveal for the first time that 25(OH)D exerts antitumor effects by promoting exon 13 skipping of KDM6A (KDM6A Δexon13), which suppresses the proliferation and stemness of breast cancer cells and lacks H3K27 demethylase activity. Mechanistically, CUT&Tag and RNA-seq analyses demonstrated that KDM6A Δexon13 induces the accumulation of H3K27me3 at the promoter region of TRAP1, thereby inhibiting its transcription. Consequently, the downregulation of TRAP1 reduces Smad2/3 phosphorylation. Furthermore, KHDRBS3 was identified as the splicing factor of KDM6A Δexon13 and was regulated by 25(OH)D. Notably, 25(OH)D exhibited a synergistic effect with GSK-J4, a specific inhibitor of KDM6A, in suppressing breast cancer cell growth. Collectively, our findings uncover a novel anticancer mechanism of 25(OH)D, highlight the critical role of KDM6A Δexon13 in breast cancer progression, and provide further evidence supporting the correction of 25(OH)D deficiency in breast cancer patients.

The extracellular matrix (ECM) defines the biomechanical and biochemical microenvironment of tissues, directing cell behaviour and phenotype. In the ovary, ECM must dynamically remodel in each cycle under hormonal regulation to control follicle development and produce fertilizable oocytes. Dysregulation of this process may result in aberrant formation of ECM as seen in polycystic ovary syndrome (PCOS) whose pathology includes fibrosis of the ovary and which is a major cause of infertility. PCOS is characterised by hyperandrogenism and, here, we investigate the impact of androgens on fibrosis, cell-ECM interactions and mechanosensing. We report an altered network of gene expression related to the genesis of fibrosis. Preantral follicles from C57BL/6 mice (14-15 days postpartum) were stimulated with dihydrotestosterone (DHT, 10nM) in 24/72 hours culture. Expression of fibrosis-associated genes (Eln; Ctgf; Acta2; Plod2; Hpse) significantly increased with androgen (72 h), as did TGF-β signalling (Tgfb1; Tgfb3). We show a direct connection between androgen and mechanosensing within the ovary, with androgen upregulating the mechanosensitive Hippo pathway (Yap1; Lats1; Lats2; Stk3; Stk4; Frmd6) and downstream targets (Ctgf; Axl; Cyr61). Our results highlight hyperandrogenism as a probable driver of the fibrosis in the polycystic ovary, and emphasise the importance of ECM regulation in follicle development and fertility.

Serine metabolism is a critical vulnerability in cancer; however, its role in mediating therapeutic resistance in non-small cell lung cancer (NSCLC) remains incompletely understood. In this study, we identify key enzymes in the serine synthesis pathway (SSP), namely PHGDH, PSAT1 and PSPH, as well as the serine transporter SLC1A4, which are significantly overexpressed in lung cancer and correlate with poor patient prognosis. We show that serine contributes to carboplatin resistance in NSCLC, particularly in lung squamous cell carcinoma (LUSC). Notably, the LUSC lineage-specific oncogene ΔNp63α serves as a master transcriptional regulator of serine biosynthesis, directly transactivating the expression of PHGDH, PSAT1, PSPH, and SLC1A4. ΔNp63α-driven serine biosynthesis supports nucleotide synthesis and enhances antioxidant defense, enabling cancer cells to survive carboplatin-induced DNA damage and oxidative stress, thereby promoting therapeutic resistance. The combined inhibition of endogenous serine synthesis and restriction of exogenous serine/glycine significantly overcomes ΔNp63α-mediated carboplatin resistance. Our findings establish the ΔNp63α-SSP axis as a critical mechanism driving carboplatin resistance in LUSC. These results highlight dual-targeted disruption of serine availability as a promising therapeutic strategy to overcome chemotherapy resistance in ΔNp63α-driven LUSC. This study underscores the importance of lineage-specific metabolic dependencies as essential targets for precision oncology in NSCLC.

Interferon tau (IFNT), a pregnancy recognition factor in ruminants, maintains the corpus luteum and induces the expression of many genes in the uterus; however, the functions of these genes remain unclear. In this study, we investigated the effect of IFNT on endothelial cell-specific molecule 1 (ESM1) expression, along with the subsequent roles of ESM1 in bovine vascular endothelial cells in an in vitro cell culture environment. Quantitative polymerase chain reaction (qPCR) analysis showed that IFNT treatment significantly increased the expression levels of ESM1 in endometrial epithelial cells compared with those in non-treated control cells. qPCR and Western blot analyses showed an increasing trend in vascular endothelial growth factor (VEGF) expression following ESM1 treatment in bovine vascular endothelial cells. In addition, ESM1 treatment tended to increase cell proliferation and significantly increased the formation of tube-like structures of endothelial cells compared to those in non-treated control cells. These results suggest that IFNT induces ESM1 expression in endometrial epithelial cells. Moreover, ESM1 may contribute to angiogenesis via the expression of angiogenic factors, such as VEGF, in bovine endothelial cells.

Brassica napus, a major oilseed crop in southern China, faces dual challenges of cadmium (Cd(II)) contamination and potassium (K) deficiency in soils. However, the mechanisms by which K alleviates Cd(II) toxicity in B. napus remain poorly understood. This study, therefore, aimed to elucidate the physiological, biochemical, and molecular mechanisms underlying K-mediated Cd(II) detoxification in B. napus using a hydroponic experimental system. The results demonstrated that elevated K application significantly alleviated Cd(II)-induced toxicity. A major finding was the substantial recovery of photosynthetic activity, with K supplementation increasing the photosynthetic rate by 40.7 % under Cd(II) stress, which corresponded with a significant reduction in biomass loss and decreased concentrations of malondialdehyde and H2O2. Mechanistically, this alleviation was linked to a fortified antioxidant defense system, evidenced by enhanced activities of catalase (CAT) and peroxidase (POD) by 54.2 % and 15.7 %, respectively. Transcriptomic analysis provided deeper insights, revealing that K-mediated tolerance involves enhanced Cd(II) sequestration. K supplementation significantly up-regulated genes encoding tonoplast-localized transporters (CAX and HMA) responsible for vacuolar Cd(II) import, while simultaneously down-regulating the vacuolar Cd(II) efflux gene NRAMP4. This was complemented by an increase in ion-bound pectin and cellulose in the cell wall, further restricting Cd(II) mobility. Furthermore, K appeared to modulate key signaling pathways, as the expression of PYL genes, encoding abscisic acid (ABA) receptors, was significantly induced. Collectively, our findings demonstrate that K alleviates Cd(II) toxicity in B. napus through a multi-pronged strategy encompassing the enhancement of antioxidant capacity and the promotion of Cd(II) sequestration in both vacuoles and cell walls, potentially regulated via an ABA-dependent pathway. This research provides a theoretical basis for optimizing K fertilization to improve phytoremediation efficiency in Cd-contaminated agricultural soils.

Low temperature stress is a major abiotic constraint on agricultural productivity, especially in temperature-sensitive crops like lettuce. Nano-selenium has demonstrated considerable potential in improving plant stress resilience. In this study, lettuce plants exposed to low-temperature stress were treated with five concentrations (N1: 1 mg L-1; N2: 3 mg L-1; N3: 9 mg L-1; N4: 27 mg L-1) of nano-selenium. The optimal concentration of nano-selenium was determined to be N3 (9 mg L-1). Integrated transcriptomic and metabolomic analyses revealed that nano-selenium application significantly enhanced photosynthetic efficiency, antioxidant defenses, and metabolic adaptation under cold stress. A total of 25,593 differentially expressed genes (DEGs) and 20 key metabolites were identified. Enriched metabolic pathways included arginine and proline metabolism, amino sugar and nucleotide sugar metabolism, and glycerophospholipid metabolism. Under low-temperature conditions, nano-selenium treatment markedly improved cold tolerance by modulating proline metabolism-promoting its biosynthesis while inhibiting its catabolism-resulting in substantial proline accumulation. Furthermore, nano-selenhanced cellular structural integrity through two distinct mechanisms: (1) reinforcing cell wall architecture via enhanced amino sugar metabolism, thereby mitigating low-temperature-induced membrane damage; and (2) optimizing glycerophospholipid composition, particularly by regulating phosphatidylcholine and phosphatidylethanolamine biosynthesis through key enzyme modulation, which helped maintain membrane fluidity and stability under cold stress. These findings advance our understanding of nano-selenium-mediated stress tolerance and underscore its potential application in sustainable agriculture.

Methotrexate (MTX) is used in treating several malignancies. However, MTX neurotoxicity remains a significant clinical side effect, leading to cell division malformation, and neurogenesis impairment. Chrysin, a flavonoid compound found in natural products, demonstrates various biological characteristics, including neuroprotective and antioxidant properties. The purpose of this study was to investigate the ameliorative effect of chrysin on oxidative damage and neurogenesis impairment caused by MTX. Male Sprague-Dawley rats were randomly divided into four groups, including the vehicle, MTX (75 mg/kg), chrysin (10 mg/kg), and chrysin+MTX groups. Chrysin was orally administered for 15 days. MTX was administered intravenously on days 8 and 15. The hippocampal neural stem cells were evaluated using sex determining region Y-box 2 (sox2) and nestin immunofluorescence staining. Antioxidant enzyme expression and the levels of oxidative stress marker were assessed. Additionally, the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), brain-derived neurotrophic factor (BDNF), cAMP-response element binding (CREB), and phosphorylated CREB (pCREB) were evaluated using Western blotting. Results showed that MTX significantly decreased the activity of antioxidant enzymes and produced oxidative stress. MTX also impaired neurogenesis, evidenced by decreased sox2 and nestin-positive cells and decreased expression of Nrf2, BDNF, CREB, and pCREB in the hippocampus and prefrontal cortex. However, chrysin significantly reversed the effects of MTX on these parameters. In conclusion, chrysin exhibits neuroprotective effects against MTX-induced neurogenesis impairment by upregulating antioxidant enzyme activity, reducing oxidative stress, and improving protein expression related to neurogenesis.

Grapevine Trunk Diseases (GTDs) are caused by phytopathogenic fungi that compromise grapevine productivity and wine quality. Most GTDs preventive treatments are chemical-based and environmentally harmful. One goal of the European Green Deal is to develop sustainable agriculture which does not harm the environment and reduces pesticide use and an alternative to those treatments may be the use of elicitors such as oligosaccharides from fungi. Many studies confirm that oligosaccharides activate the defence response. The experiment was carried out in vineyards of Tempranillo and Airén cvs. Asymptomatic and symptomatic vines were treated with mannans. Leaves and grapes were taken and pigments and phenols content, polyphenol oxidase (PPO) and superoxide dismutase (SOD) activities and gene expression of several defence enzymes were determined. The mannan addition to symptomatic vines was more positive for the leaves than for the grapes, palliating the damage caused by the disease, especially in the cv. Tempranillo. On the one hand, in the leaves, mannans caused an increase in phenols and PPO activity and expression; on the other hand, in grapes, although phenols increased, the other parameters did not. Mannans increased the expression levels of chalcone synthase (CHS1, CHS3), phenylalanine ammonia lyase (PAL), SOD, and PPO in asymptomatic leaves of both cultivars. In symptomatic leaves, CHS3 and PAL expression decreased in both cultivars, while CHS1 and PPO increased only in Tempranillo. In grapes, the expression of the genes varied due to the development of the disease. The mannan treatment seemed to reduce the oxidative stress caused by GTDs, but, above all, mannans would act as a biostimulant activaing the defence system of asymptomatic vines that would help them respond more successfully to a possible pathogenic fungi infection, that although this response depended on the cultivar.

The fungal priority pathogen and basidiomycete, Cryptococcus neoformans (Cn), causes lung and brain infection in predominantly immuno-compromised individuals and there is an urgent need for new treatment options. The pyrazolopyrimidine-based cyclin dependent kinase (CDK)7 inhibitor, BS-181, has anticancer properties, but its antifungal activity has not been investigated. We show that cryptococcal CDK7 more closely resembles the human enzyme than that of ascomycetes, and that BS-181 inhibits its activity. BS-181 inhibited growth of both Cn and Cryptococcus gattii (Cg), but not ascomycete fungi and delayed progression through the G2/M phase of the cell cycle. Transcriptomic analysis revealed that BS-181 induces splicing defects leading to elevated intron retention within the transcriptome and also suppresses translational processes. BS-181 displayed additive or synergistic activity with licensed antifungals against laboratory and clinical Cn and Cg strains, most notably with amphotericin B where synergy (2-4-fold reduction in the amphotericin B MIC) was achieved using low-sub micromolar concentrations of BS-181. Compared with either drug alone, BS-181-AmB combination therapy provided greater protection against Cn infection in a wax moth model (p ≤ 0.032) and extended survival of Cn-infected mice. These findings demonstrate that CDK7 inhibitors, already of interest as anticancer agents, could be repurposed to prevent or treat opportunistic fungal infections in cancer patients when combined with licensed antifungals limited by either toxicity or resistance.

IntroductionGlycolytic phenotype positively supports cancer cell migration and metastasis in various cancers including Triple negative breast cancers (TNBCs). In-depth understanding of molecular pathways associated with increased aerobic glycolysis in TNBCs could provide key insights into the drivers of TNBC progression.Methodsβ-catenin and glycolytic proteins (PFKP, LDHA, MCT1) were assessed by Immunohistochemistry (IHC) in TNBC patients (n = 98), with prognostic value evaluated by Kaplan-Meier and Cox regression. In vitro, the β-catenin inhibitor ie, XAV939 was tested for suppressing β-catenin-driven aerobic glycolysis in TNBC models using MTT for proliferation, Western blotting for protein expression, and wound healing, droplet invasion, and colony formation assays for physiological changes.Resultsβ-catenin and glycolytic markers (PFKP, LDHA, MCT1) were overexpressed in >50% of TNBCs. Kaplan-Meier and Cox regression analyses showed that combined expression of β-catenin with glycolytic markers correlated with reduced survival. In vitro, XAV939 suppressed β-catenin-driven aerobic glycolysis in TNBC cells, downregulating β-catenin and glycolytic proteins, reducing glycolytic activity, and impairing aggressive phenotypes (proliferation, migration, invasion, clonogenicity).ConclusionOverall, our results highlight the crucial role of β-catenin in controlling aerobic glycolysis via regulation of key glycolytic proteins, thereby positively driving the progression and metastasis of TNBCs. Additionally, our data strongly establish that XAV939 effectively inhibits glycolytic phenotype, thereby suggesting its therapeutic potential in TNBC patients.

Clinical use of stimulator of interferon genes (STING) agonists has challenges due to poor responsiveness and variable efficacy. Therefore, identifying tumor types that are sensitive to these agents and clarifying the underlying mechanisms are essential.

Liver cancer treatment with cisplatin is often hindered by drug resistance. This study aimed to identify key genes associated with cisplatin resistance in liver cancer and develop targeted inhibitors. Using genome-wide CRISPR-Cas9 screening, ATOX1 was identified as a critical gene for cisplatin resistance. ATOX1 was highly expressed in liver cancer tissues and associated with poor prognosis. Knockdown of ATOX1 in liver cancer cells enhanced cisplatin sensitivity in vitro and in vivo. Molecular dynamics simulation and virtual screening identified compound 8 as a potent ATOX1 inhibitor with high affinity (Kd = 12.5 μM) and exhibited synergistic effects with cisplatin on liver cancer cell growth. Mechanistically, compound 8 inhibits the activity of ATOX1, leading to intracellular copper accumulation. The elevated copper levels subsequently promote increased DNA methylation at the NOTCH1 promoter, resulting in suppression of the NOTCH1/HES1 signaling pathway and enhancing the sensitivity of liver cancer cells to cisplatin. In conclusion, ATOX1 is crucial for cisplatin resistance in liver cancer and linked to poor prognosis. Targeting ATOX1 with compound 8 may be a novel therapeutic strategy for overcoming cisplatin resistance.

This study assesses the neuroprotective effects and molecular mechanisms of ciprofol against cerebral ischemia-reperfusion injury (CIRI) in rats. From July 2023 and July 2024, fifty male SD rats were randomly divided into five groups: control, model, ciprofol (Ci), erastin (Era), and compound C (CC). The model was induced by MCAO/R. Control group received identical surgical interventions without filament insertion. Rats neurological deficits were quantified using modified Garcia JH scores. Histopathological changes were evaluated through Nissl staining and TTC‒determined infarct volume. Mitochondrial ultrastructure was observed by transmission electron microscopy. Biochemical analyses quantified malondialdehyde (MDA), iron content and inflammatory cytokines (IL-1β, IL-6, TNF-α). Western blotting measured AMPK phosphorylation and ferroptosis proteins (GPX4, ACSL4). Relative to the model group rats, the Ci rats exhibited elevated modified Garcia JH scores (P < 0.05) accompanied by attenuated neuronal/mitochondrial damage and diminished infarct areas. Biochemical analyses revealed significant reductions in MDA, inflammatory cytokines, and ACSL4 protein expression in Ci rats, concurrent with enhanced GPX4 levels and elevated p-AMPK/T-AMPK ratios (P < 0.05). Notably, Era specimens demonstrated GPX4 downregulation with corresponding ACSL4 and iron accumulation compared to the Ci rats. Furthermore, the CC rats displayed decreased p-AMPK/T-AMPK activation relative to Ci rats (P < 0.05). Ciprofol ameliorates CIRI in rats by inhibiting ferroptosis and inflammatory factor through upregulating AMPK.

Although many DNA binding natural products exert their effects through non-specific mechanisms, a therapeutic opportunity exists for a subset of these compounds that alter the expression or activity of specific driver oncogenes in specific cell contexts. In this study, we integrate CUT&Tag with Global Run-On Sequencing (CUT, Tag, and GRO) to show that the minor groove binding compound, mithramycin (MMA), inhibits the Ewing sarcoma oncogenic driver, the EWS::FLI1 transcription factor. MMA causes either an increase or decrease in EWS::FLI1 binding to chromatin at downstream target response elements to poison nascent transcription. The reversal of EWS::FLI1 activity is limited by non-specific effects of the drug on RNAPII processivity but can be optimized by continuous administration at low concentration to cause more precise reversal of the oncogenic transcriptome and striking Ewing sarcoma xenograft regressions. The activity in vivo is further improved with a less-toxic second-generation analog, AIT-102.

BackgroundHypopharyngeal cancer is increasingly emerging as a disease that threatens global health, with poor prognosis and survival rates. However, clinical strategies and effective therapies remain limited.MethodsThe inhibitory effect of liensinine on tumor cells was detected through cell cycle, colony formation, and apoptosis assays. Changes in the expression levels of relevant proteins were detected and enrichment analysis of signaling pathways was performed through in vitro and RNA sequencing experiments. The transcription levels of relevant genes were further verified using reverse transcription polymerase chain reaction.ResultsWe previously discovered that the natural compound, liensinine, is effective in treating hypopharyngeal cancer. In this study, we found through in vitro and RNA sequencing experiments that liensinine can activate the Ras homolog family member B protein, thereby inhibiting the mitogen-activated protein kinase signaling pathway. Additionally, liensinine activates the nuclear factor kappa B signaling pathway and releases downstream inflammatory factors, effectively exerting its antitumor effects.ConclusionLiensinine induces cell death and inhibits hypopharyngeal cancer cell growth through multiple pathways, indicating that it is a potential chemotherapeutic agent for the treatment of hypopharyngeal cancer.

High myopia (HM) poses a growing public health challenge due to its increasing prevalence and the associated risks of blinding complications and psychological comorbidities. While traditionally considered an isolated ocular condition, emerging evidence implicates systemic mechanisms, notably through the gut-eye axis and immune factors, play important part in the pathogenesis of HM. Histone demethylase Kdm2a, the key H3K36me2 modification eraser, is critically involved in various inflammatory diseases, yet its specific role in the gut-eye axis and HM remains elusive. To address this, the HM model was successfully established. HM mice exhibited significant scleral thinning, reduced collagen protein, and prominent anxiety-like behaviors. Crucially, they were suffering from gut microbial dysbiosis and intestinal barrier impairment. Intriguingly, upregulated Kdm2a and correspondingly decreased H3K36me2 levels were observed in the intestinal epithelial cells (IECs) of HM mice. Treatment with Daminozide (DA), the selective inhibitor of Kdm2a, effectively suppressed myopia progression and ameliorated psychological comorbidities. Mechanistically, DA restored gut microbiota homeostasis, colonic morphology, and barrier integrity. The transcriptomic profiling further revealed the protective effects of Kdm2a inhibition on modulating key pathways involved in intestinal inflammation and tissue remodeling. Collectively, this work elucidates a novel gut-eye pathway in HM pathogenesis and identifies Kdm2a in IECs as a promising therapeutic target for HM and its associated psychological comorbidities.

Intracerebral hemorrhage (ICH) is a cerebrovascular event associated with a high fatality rate, leading to a considerable health and economic burden. Tanshinone IIA (Tan IIA), a promising compound used to treat coronary artery disease, has recently been shown to exert significant neuroprotective effects. Therefore, whether Tan IIA can alleviate NETosis induced by LPS after ICH remains unclear. For this purpose, we explored the effects of Tan IIA on collagenase-induced ICH with peripheral inflammation and its potential mechanisms using an after-ICH infection animal model (male C57BL/6J mice) treated with Tan IIA for 5 days, starting at 2 months of age. Further analysis demonstrated that Tan IIA-treated ICH mice with peripheral inflammation exhibited improved motor and sensory dysfunction compared with untreated groups. Administration of Tan IIA in ICH mice with peripheral inflammation alleviated neuropathological alterations of the corpus striatum, including NETosis inhibition, glial inactivation, and inflammasome activity attenuation, and significantly decreased levels of PAD4 and H3 Cit in the corpus striatum of ICH mice with peripheral inflammation. In vitro investigations showed that Tan IIA suppressed neuroinflammation in LPS-stimulated glial cells by inhibiting the NLRP3/caspase-1 signaling pathway. Further molecular docking predicted that Tan IIA directly interacted with the NLRP3 protein. Collectively, these findings strongly indicate that Tan IIA is an effective compound for mitigating hemiplegia symptoms, NETosis, and neuroinflammation in the collagenase-induced ICH model with peripheral inflammation, primarily through the dual actions of inhibiting NET formation and suppressing the NLRP3/caspase-1 pathway.

Isolavonoides represent the second largest subgroup of flavonoids and have an influence on critical molecular pathways and restore cellular homeostasis, through the reprogramming of epigenetic regulatory mechanisms. This feature indicates a crucial therapeutic potential that could be better explored to attend cancer treatment. Isoflavonoids, acting as epigenetic modulators, could contribute to the development of new therapeutic approaches in cancer, especially in onco-hematological diseases. Pterocarpans are a subgroup of isoflavonoids that have been extensively studied for their biological properties. The molecule (+)-2,3,9-trimethoxypterocarpan demonstrates high gastrointestinal (GI) absorption and the ability to cross the blood-brain barrier (BBB) in silico without violating Lipinski's rule, making it a desirable candidate in leukemia treatment. The synthesis of this molecule dates back more than a decade. In silico models, such as SwissADME, corroborate the notion of good intestinal absorption and the ability to cross the BBB. Also, it is suggested that P-glycoprotein is a substrate, which is related to its potential for active efflux from both the BBB and GI. This review highlights the biological mechanisms of this class of natural products from a translational perspective, emphasizing their chemical properties and epigenetic biological activities, which offer new therapeutic perspectives, particularly in oncology.

Jasmonate (JA) accumulation and signaling play key roles in regulating plant growth and flower development. The optimal timing of flowering is critical for the quality and yield of Brassica juncea. B-box (BBX) factors play roles in plant floral transition and stress response processes. However, whether BBX6-2 responds to JA signaling to regulate flowering time in B. juncea remains unclear. Here, we characterized the biological function of BjuBBX6-2 in response to JA in regulating the flowering time of B. juncea. Subcellular localization and transcriptional activation activity assays showed that BjuBBX6-2 localizes in the nucleus and exhibits transcriptional activation activity. Spraying BjuBBX6-2-overexpressing or BjuBBX6-2-silenced B. juncea plants with 50 μmol/L methyl JA significantly accelerated or delayed their flowering time, respectively. We demonstrated that BjuBBX6-2 interacts with the flowering factor Nuclear Factor Y, Subunit C4 (BjuNF-YC4), which interacts with Nuclear Factor Y, Subunit B2/3 (BjuNF-YB2/3). The BjuBBX6-2-BjuNF-YC4-BjuNF-YB2/3 multiple-protein complex bound to the promoter of the downstream flowering integrator gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (BjuSOC1) and promoted its expression. JASMONATE ZIM-DOMAIN 1 (BjuJAZ1), a key factor in the JA signaling pathway, interacted with BjuBBX6-2 to inhibit the activation of BjuSOC1 by BjuBBX6-2. In summary, BjuBBX6-2 cooperates with BjuNF-YC4 and BjuJAZ1 in response to JA signaling to participate in the flowering regulation of B. juncea. These findings highlight a previously uncharacterized mechanism of JA signaling-mediated flowering time regulation via interactions between BjuBBX6-2 and the integrator gene BjuSOC1, providing prospects for breeding enhanced B. juncea cultivars.